目的制备一种结合链霉亲和素的高分子超声造影剂,并与生物素化抗体结合,考察其体外寻靶能力。方法采用双乳化法制备高分子材料PLGA-COOH超声造影剂;并用碳二亚胺法将造影剂与亲和素耦联,制备出结合链霉亲和素的高分子超声造影剂。检测该造影剂一般特性;采用红外与免疫荧光检测亲和素与PLGA-COOH超声造影剂连接的情况。检测生物素-亲和素技术使该造影剂与生物素化抗体结合后其体外寻靶能力。结果红外检测与免疫荧光检测显示亲和素被成功地连接在高分子造影剂上,并存在活性。体外寻靶实验显示,与生物素化单抗结合后的靶向高分子造影剂较多并牢固地聚集到肝癌细胞表面。结论成功制备了结合链霉亲和素的高分子超声造影剂,该造影剂与生物素化抗体结合后于体外对肝癌细胞具有较强的亲和力。
Objective To prepare a new kind of high molecular polymer ultrasound contrast agents integrated with streptoavidin,which were combined with the biotinylated antibody,and to investigate their ability to find target in vitro.Methods The high molecular polymer ultrasound contrast agents integrated with streptoavidin were produced using the technique of double emulsion and EDC/NHS.Besides the usual characteristics,the connection between the high molecular polymer ultrasound contrast agent and streptoavidin was detected with methods of infrared spectra and immunofluorescence.The targeting ability of the contrast agent combined with biotinylated antibody was contrasted with that of non-combined in vitro.Results Infrared spectra and immunofluorescent assay showed that streptoavidin was successfully conjugated on the surface of polymer contrast agent with activity.In vitro,many targeted high molecular polymer ultrasound contrast agents could conjugate with hepatocarcinoma tightly.Conclusion This high molecular polymer ultrasound contrast agents are prepared conjugated with streptoavidin successfully,which can target to hepatocarcinoma effectively in vitro through combination with the biotinylated antibody.